Colorimetric analysis for measuring activity of cellular enzymes that reduce a tetrazolium dye
A microtiter plate after an MTT assay. Increasing amounts of cells resulted in increased purple colouring.
The MTT assay is a colorimetric assay for assessing cell metabolic activity.[1][2] NAD(P)H-dependent cellular oxidoreductase enzymes may, under defined conditions, reflect the number of viable cells present. These enzymes are capable of reducing the tetrazolium dye MTT, which is chemically 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide, to its insoluble formazan, which has a purple color. Other closely related tetrazolium dyes including XTT, MTS and the WSTs, are used in conjunction with the intermediate electron acceptor, 1-methoxy phenazine methosulfate (PMS). With WST-1, which is cell-impermeable, reduction occurs outside the cell via plasma membrane electron transport.[3] However, this traditionally assumed explanation is currently contended as proof has also been found of MTT reduction to formazan in lipidic cellular structures without apparent involvement of oxidoreductases.[4]
Tetrazolium dye assays can also be used to measure cytotoxicity (loss of viable cells) or cytostatic activity (shift from proliferation to quiescence) of potential medicinal agents and toxic materials. MTT assays are usually done in the dark since the MTT reagent is sensitive to light.[5][6][7]
^Gavanji S, Bakhtari A, Famurewa AC, Othman EM (January 2023). "Cytotoxic Activity of Herbal Medicines as Assessed in Vitro: A Review". Chemistry & Biodiversity. 20 (2): 3–27. doi:10.1002/cbdv.202201098. PMID 36595710. S2CID 255473013.
^Stockert JC, Horobin RW, Colombo LL, Blázquez-Castro A (April 2018). "Tetrazolium salts and formazan products in Cell Biology: Viability assessment, fluorescence imaging, and labeling perspectives" (PDF). Acta Histochemica. 120 (3): 159–167. doi:10.1016/j.acthis.2018.02.005. PMID 29496266. Archived (PDF) from the original on 2022-02-24. Retrieved 2020-04-15.
^Berridge MV, Herst PM, Tan AS (2005). "Tetrazolium dyes as tools in cell biology: new insights into their cellular reduction". Biotechnology Annual Review. 11: 127–152. doi:10.1016/S1387-2656(05)11004-7. ISBN 9780444519528. PMID 16216776.
^Stockert JC, Blázquez-Castro A, Cañete M, Horobin RW, Villanueva A (December 2012). "MTT assay for cell viability: Intracellular localization of the formazan product is in lipid droplets". Acta Histochemica. 114 (8): 785–796. doi:10.1016/j.acthis.2012.01.006. PMID 22341561.
^Sharma, Dr Rudra (2022-10-05). "Cytotoxicity Test by MTT Assay- Procedure, Analysis and Results - ACME Research Solutions". Retrieved 2022-10-06.
^Müller, G.; Kramer, A. (2006). "Comparative Study of in vitro Cytotoxicity of Povidone-Iodine in Solution, in Ointment or in a Liposomal Formulation (Repithel<sup>®</sup>) and Selected Antiseptics". Dermatology. 212 (Suppl. 1): 91–93. doi:10.1159/000090102. ISSN 1018-8665. PMID 16490982. S2CID 818013.
^Pintor, A. V. B.; Queiroz, L. D.; Barcelos, R.; Primo, L. S. G.; Maia, L. C.; Alves, G. G. (2020-07-27). "MTT versus other cell viability assays to evaluate the biocompatibility of root canal filling materials: a systematic review". International Endodontic Journal. 53 (10): 1348–1373. doi:10.1111/iej.13353. ISSN 0143-2885. PMID 32602945. S2CID 220271896.
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